TY - JOUR

T1 - Viral Strategies for Targeting Spinal Neuronal Subtypes in Adult Wild-Type Rodents

AU - Kaur, Jaspreet

AU - Berg, Rune W.

N1 - Targeting specific subtypes of interneurons in the spinal cord is primarily restricted to a small group of genetic model animals. Since the development of new transgenic model animals can be expensive and labor intensive, it is often difficult to generalize these findings and verify them in other model organisms, such as the rat, ferret or monkey, that may be more beneficial in certain experimental investigations. Nevertheless, endogenous enhancers and promoters delivered using an adeno-associated virus (AAV) have been successful in providing expression in specific subtypes of neurons in the forebrain of wildtype animals, and therefore may introduce a shortcut. GABAergic interneurons, for instance, have successfully been targeted using the mDlx promotor, which has recently been developed and is now widely used in wild type animals. Here, we test the specificity and efficiency of the mDlx enhancer for robust targeting of inhibitory interneurons in the lumbar spinal cord of wild-type rats. Since this has rarely been done in the spinal cord, we also test the expression and specificity of the CamKIIa and hSynapsin promoters. We found that mDlx does in fact target many neurons that contain an enzyme for catalyzing GABA, the GAD-65, with high specificity and a small fraction of neurons containing an isoform, GAD-67. Expression was also seen in some motor neurons. Viral injections using the CamKIIa enhancer infected in some glutamatergic neurons but also GABAergic neurons, whereas hSynapsin targets almost all the neurons in the lumbar spinal cord.

PY - 2022

Y1 - 2022

N2 - Targeting specific subtypes of interneurons in the spinal cord is primarily restricted to a small group of genetic model animals. Since the development of new transgenic model animals can be expensive and labor intensive, it is often difficult to generalize these findings and verify them in other model organisms, such as the rat, ferret or monkey, that may be more beneficial in certain experimental investigations. Nevertheless, endogenous enhancers and promoters delivered using an adeno‐ associated virus (AAV) have been successful in providing expression in specific subtypes of neurons in the forebrain of wildtype animals, and therefore may introduce a shortcut. GABAergic interneurons, for instance, have successfully been targeted using the mDlx promoter, which has recently been developed and is now widely used in wild type animals. Here, we test the specificity and efficiency of the mDlx enhancer for robust targeting of inhibitory interneurons in the lumbar spinal cord of wild‐type rats using AAV serotype 2 (AAV2). Since this has rarely been done in the spinal cord, we also test the expression and specificity of the CamKIIa and hSynapsin promoters using serotype 9. We found that AAV2-mDlx does in fact target many neurons that contain an enzyme for catalyzing GABA, the GAD‐65, with high specificity and a small fraction of neurons containing an isoform, GAD‐67. Expression was also seen in some motor neurons although with low correlation. Viral injections using the CamKIIa enhancer via AAV9 infected in some glutamatergic neurons, but also GABAergic neurons, whereas hSynapsin via AAV9 targets almost all the neurons in the lumbar spinal cord.

AB - Targeting specific subtypes of interneurons in the spinal cord is primarily restricted to a small group of genetic model animals. Since the development of new transgenic model animals can be expensive and labor intensive, it is often difficult to generalize these findings and verify them in other model organisms, such as the rat, ferret or monkey, that may be more beneficial in certain experimental investigations. Nevertheless, endogenous enhancers and promoters delivered using an adeno‐ associated virus (AAV) have been successful in providing expression in specific subtypes of neurons in the forebrain of wildtype animals, and therefore may introduce a shortcut. GABAergic interneurons, for instance, have successfully been targeted using the mDlx promoter, which has recently been developed and is now widely used in wild type animals. Here, we test the specificity and efficiency of the mDlx enhancer for robust targeting of inhibitory interneurons in the lumbar spinal cord of wild‐type rats using AAV serotype 2 (AAV2). Since this has rarely been done in the spinal cord, we also test the expression and specificity of the CamKIIa and hSynapsin promoters using serotype 9. We found that AAV2-mDlx does in fact target many neurons that contain an enzyme for catalyzing GABA, the GAD‐65, with high specificity and a small fraction of neurons containing an isoform, GAD‐67. Expression was also seen in some motor neurons although with low correlation. Viral injections using the CamKIIa enhancer via AAV9 infected in some glutamatergic neurons, but also GABAergic neurons, whereas hSynapsin via AAV9 targets almost all the neurons in the lumbar spinal cord.

KW - Faculty of Health and Medical Sciences

KW - spinal cord

KW - AAV virus

KW - Rodent

U2 - 10.1038/s41598-022-12535-4

DO - 10.1038/s41598-022-12535-4

M3 - Journal article

C2 - 35606530

VL - 12

JO - Scientific Reports

JF - Scientific Reports

SN - 2045-2322

IS - 1

M1 - 8627

ER -